Activation of TNFα pathways. A. Detection of TNFα by ELISA in the neural retina. Cytokine levels were determined in three samples at each time point. The level at 6 hr was significantly higher. B-C. Detection of phosphorylated JNK in the neural retina. B. Western blots (upper image) were probed with an anti pJNK antibody. C. ELISA confirmed the change in activation of JNK at 6 hrs after optic nerve injury. Data are normalized to total protein. D-G. Immunohistochemistry of activated JNK in the neural retina. Antibodies to pJNK were used on sections at 0, 30 min, 60 min and 6 hr after optic nerve crush. Increased labeling is evident in the GCL (arrows) at 6 hr.